Micro-Pillar Array Column Separations for Proteomics of Liver Organoids

Organoids are laboratory-grown three-dimensional (3D) models of organs and are emerging tools for studies into developmental biology, drug discovery, and toxicology. Organoids are complex biological materials, and proteomics studies of organoids can benefit from applying high-resolution chromatography devices before mass spectrometric analysis. Micro-pillar array columns have been shown to provide excellent resolution of peptide products of proteolytic digestion of proteins for bottom-up proteomics.This article describes a workflow incorporating a micro-pillar array column for mapping the proteome of human stem cell-derived liver organoids (sample preparation using a SPEED protocol) using trapped ion mobility time-of-flight mass spectrometry (timsTOF-MS).